Active optics such as deformable mirrors can be used to control both focal depth and aberrations during scanning laser microscopy. If the focal depth can be changed dynamically during scanning, then imaging of oblique surfaces becomes possible. If aberrations can be corrected dynamically during scanning, an image can be optimized throughout the field of view. Here, we characterize the speed and dynamic precision of a Boston Micromachines Corporation Multi-DM 140 element aberration correction mirror and a Revibro Optics 4-zone focus control mirror to assess suitability for use in an active and adaptive two-photon microscope. Tests for the multi-DM include both step response and sinusoidal frequency sweeps of specific Zernike modes (defocus, spherical aberration, coma, astigmatism, and trefoil). We find wavefront error settling times for mode amplitude steps as large as 400 nm to be less than 52 μs, with 3 dB frequencies ranging from 6.5 to 10 kHz. The Revibro Optics mirror was tested for step response only, with wavefront error settling time less than 80 μs for defocus steps up to 3000 nm, and less than 45 μs for spherical aberration steps up to 600 nm. These response speeds are sufficient for intrascan correction at scan rates typical of two-photon microscopy.
Active optics can facilitate two-photon microscopic imaging deep in tissue. We are investigating fast focus control mirrors used in concert with an aberration correction mirror to control the axial position of focus and system aberrations dynamically during scanning. With an adaptive training step, sample-induced aberrations may be compensated as well. If sufficiently fast and precise, active optics may be able to compensate under-corrected imaging optics as well as sample aberrations to maintain diffraction-limited performance throughout the field of view. Toward this end we have measured a Boston Micromachines Corporation Multi-DM 140 element deformable mirror, and a Revibro Optics electrostatic 4-zone focus control mirror to characterize dynamic performance. Tests for the Multi-DM included both step response and sinusoidal frequency sweeps of specific Zernike modes. For the step response we measured 10%-90% rise times for the target Zernike amplitude, and wavefront rms error settling times. Frequency sweeps identified the 3dB bandwidth of the mirror when attempting to follow a sinusoidal amplitude trajectory for a specific Zernike mode. For five tested Zernike modes (defocus, spherical aberration, coma, astigmatism and trefoil) we find error settling times for mode amplitudes up to 400nm to be less than 52 us, and 3 dB frequencies range from 6.5 kHz to 10 kHz. The Revibro Optics mirror was tested for step response only, with error settling time of 80 μs for a large 3 um defocus step, and settling time of only 18 μs for a 400nm spherical aberration step. These response speeds are sufficient for intra-scan correction at scan rates typical of two-photon microscopy.